Studies of Antibody Diversity and Genome Stability: Regulation of Activation-Indu

抗体多样性和基因组稳定性的研究：激活诱导的调节

• 批准号: 8002358
• 负责人: Yee Ling Wu
• 金额: $ 3.98万
• 依托单位: MEDICAL RES COUNCIL LAB OF MOLEC BIOL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-11-24 至 2013-11-23
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8002358
• 关键词: Antibodies; Antibody Diversity; Biochemical; Cancer Etiology; Cell physiology; DNA; DNA Sequence; Disease; Enzymes; Fellowship; Generation of Antibody Diversity; Genes; Genetic Transcription; Genome Stability; Genomics; Goals; Hereditary Disease; Immune; Immunologics; Infection; Knowledge; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Microbe; Molecular; Mutate; Mutation; Normal Cell; Organism; Physiological; Proteins; RNA Splicing; Regulation; Vertebrates; activation-induced cytidine deaminase; immune function; leukemia/lymphoma; microbial; next generation; novel therapeutics; public health relevance; research study

DESCRIPTION (provided by applicant): Activation-induced deaminase (AID) is an enzyme that mutates genes for antibodies in vertebrates. These AID-mediated mutations are necessary for an organism to generate specific antibodies to recognize and eradicate great varieties of microbial infections. However, if AID is not properly regulated, it can change DNA sequences of genes important for normal cell functions and cause diseases. Indeed, some lymphomas and leukemias are caused by abnormal activities of AID. How AID can change DNA sequences only in antibody genes but not in other genes is poorly understood. Recently, the catenin 2 like 1 protein (CTNNBL1) has been found to physically interact with AID. Little is known for the physiologic function of CTNNBL1. However, the interaction with CTNNBL1 is required for AID to change DNA sequences specifically in the antibody genes. In this study, a combination of high throughput approaches including next-generation sequencing and microarray analyses and targeted biochemical and immunologic analyses will be used to investigate how AID specifically mutate antibody genes, particularly what genomic regions are accessible to AID, both in the presence and in the absence of CTNNBL1. These experiments will also detect any alterations in transcription and splicing caused by the absence of CTNNBL1, and thereby shed lights on normal cellular functions of CTNNBL1. Mass spectrometry, mutagenic analyses and immunologic experiments will be performed to determine the molecular details of the interaction between AID and CTNNBL1 as well as to elucidate the mechanisms by which CTNNBL1 regulates the targeted activity of AID. It is anticipated that results from this study will be highly relevant for understanding the mechanisms underlying the generation of antibody diversity, and in the creation of new therapeutic strategies to minimize genomic disorders and cancers. PUBLIC HEALTH RELEVANCE: AID (activation-induced deaminase) is the DNA mutating enzyme responsible for changes of the antibody genes to generate different antibodies, but untimely or accidental actions of AID on cellular genes can cause cancers, immune-mediated or genetic diseases. The goal of this fellowship is to find out how AID is finely controlled to introduce mutations on genes for antibodies against microbes but not undesirable and harmful changes on genes of regular functions. Knowledge gained from this study will help understanding normal immune functions and facilitate the creation of new therapeutic strategies to minimize diseases such as cancers.

描述(申请人提供)：激活诱导脱氨酶(AID)是一种在脊椎动物中突变抗体基因的酶。这些AID介导的突变对于有机体产生识别和根除各种微生物感染的特定抗体是必要的。然而，如果AID没有得到适当的调控，它可能会改变对正常细胞功能至关重要的基因的DNA序列，并导致疾病。事实上，一些淋巴瘤和白血病是由艾滋病的异常活动引起的。AID如何只改变抗体基因的DNA序列，而不改变其他基因的DNA序列，目前还知之甚少。最近，连环蛋白2样蛋白1(CTNNBL1)被发现与AID发生物理相互作用。CTNNBL1的生理功能知之甚少。然而，AID需要与CTNNBL1相互作用才能改变抗体基因中的DNA序列。在这项研究中，包括下一代测序和微阵列分析在内的高通量方法以及有针对性的生化和免疫学分析将被用于研究AID如何特异性地突变抗体基因，特别是在存在和不存在CTNNBL1的情况下，AID可以访问哪些基因组区域。这些实验还将检测由于缺乏CTNNBL1而导致的转录和剪接的任何变化，从而揭示CTNNBL1的正常细胞功能。将通过质谱学、突变分析和免疫学实验来确定AID与CTNNBL1相互作用的分子细节，并阐明CTNNBL1调节AID靶向活性的机制。预计这项研究的结果将对理解抗体多样性产生的潜在机制以及创建新的治疗策略以最大限度地减少基因组疾病和癌症具有高度相关性。 公共卫生相关性：AID(激活诱导脱氨酶)是一种DNA突变酶，负责抗体基因的变化以产生不同的抗体，但AID对细胞基因的不及时或偶然作用可导致癌症、免疫介导或遗传性疾病。这项奖学金的目标是找出如何精细控制AID，以在针对微生物的抗体的基因上引入突变，而不是在正常功能的基因上引入不良和有害的变化。从这项研究中获得的知识将有助于了解正常的免疫功能，并有助于创造新的治疗策略，将癌症等疾病降至最低。